Nous remercions D. Ashton, V. Barkley, K. Beimfohr, F. Bojorquez, J. Cantrell, G. Castro, R. Christensen, J. Fearn, C. Jara, D. Meade, G. Owens, L. Rodarte, D. Sieglaff, A. Sonoqui, M. Stefanek, Stuart B., J. Trejo, A. Slade et Yescas E. une assistance technique. Prise en charge partielle a été fourni par USDA Agricultural Research Service, USDA-Institut National pour l’alimentation et l’Agriculture Extension IPM programme et Pest Management Solutions de rechange spécial projets, Cotton Incorporated, Arizona Association des producteurs de coton, coton Fondations, USDA-cris, NAPIAP (région de l’Ouest) et région de l’Ouest IPM projets spéciaux.

Les auteurs n’ont rien à divulguer.

En règle générale, le développement de la vie, tables pour insectes multivoltines avec largement les générations imbriquées sont limités à une approche verticale où la population est échantillonnée à plusieurs reprises dans le temps et les différentes techniques graphiques et mathématiques sont ensuite utilisés pour estimation de recrutement aux différentes étapes et déduire les taux de mortalité de changer la densité de la vie différents stades2. La force de l’approche ici...

Les tables sont un outil commun avec une longue histoire en écologie1,2. Les tables sont qu'essentiellement une planification des naissances et décès dans une population de plus de temps et ces données peut servir à mesurer un certain nombre de paramètres importants pour la compréhension et la prédiction de la dynamique des populations. Tables de survie peuvent également fournir des informations sur les causes de décès qui sont importants pour comprendre les interactions trophiques et à élaborer des stratégies de contrôle pour la gestion des ravageurs dans les systèmes agricoles et naturels. Nombreuses tables de survie sur le terrain ont été construits pour insectes3,4,5, et les analyses ont fourni des renseignements importants sur la dynamique, le règlement et la prévision des populations d’insectes dans de nombreux gérés et les systèmes naturels6,7,8,9,10,11,12,13,14. La table de survie à terme est également souvent utilisée pour décrire les études laboratoire basé en grande partie sur horaires des naissances et des décès, mais dans des conditions artificielles qui n’exposent pas l’insecte à la mortalité naturelle des forces et des variables environnementales réalistes. En règle générale, l’objectif des études en laboratoire consiste à estimer le potentiel biotique comparatif d’une espèce. La mise au point des méthodes décrites ici est pour champ basé enquêtes qui définissent réalisé le potentiel par rapport à l’environnement.
Les tables peuvent être qualifiées d’horizontal, dans lequel une cohorte réelle d’individus âgés égales sont suivies depuis le début de leur vie jusqu'à la mort, ou vertical, où les fréquentes échantillons sont prélevés à travers le temps d’une population avec une structure d’âge présumé de stable et puis vitaux est déduites des cohortes mathématiquement construit2,15. Le type de table de survie qui peut être déployée dépend de la nature de l’insecte. Tables de vie horizontale peuvent souvent être développées pour insectes univoltins (une génération par an), alors qu’une telle approche peut être très difficile pour un insecte multivoltines avec multiple et générations chaque année largement imbriquées. Une multitude de méthodes d’analyse ont été proposées et utilisées pour élaborer des tableaux de vie verticale pour les populations d’insectes (voir Southwood2 pour des exemples). La méthodologie a démontré ici permet le développement horizontal, fondées sur une cohorte des tables de survie dans le domaine des insectes multivoltines avec les caractéristiques du cycle de vie spécifique, notamment, la présence de stades sessiles. La méthode est démontrée pour un ravageur clé en coton comme système modèle.
L’aleurode, Bemisia tabaci biotype B (= Bemisia argentifolii, Moyen-Orient-Asie mineure 116) est un ravageur global de l’agriculture que les répercussions négatives sur le rendement et la qualité dans nombreuses cultures agronomiques et horticoles, y compris protégé des systèmes agricoles dans les régions tempérées,17. Les impacts se produisent en raison de phloème qui perturbe le flux des éléments nutritifs, d’étiologie inconnue, causée par l’alimentation larvaire, transmission de nombreux virus végétaux et effets de qualité de récolte en raison de la déposition du miellat18,19 , les troubles d’alimentation . L’insecte a une vaste gamme d’hôtes et multivoltines, ayant jusqu'à 12-13 générations par an selon la région et de ressources alimentaires disponibles20. Défis de gestion sont également exacerbées par son fort potentiel reproducteur, sa capacité à se disperser et de migrer au sein et entre les systèmes agricoles, son manque d’une phase de repos (diapause ou estivation) et la décision de développer rapidement une résistance aux insecticides utilisés pour répression21,22.
Des progrès considérables accomplis dans l’élaboration des stratégies de lutte antiparasitaire intégrée (LAI) pour gérer efficacement et économiquement les populations de ce ravageur dans les cultures touchées23,24,25. Ces systèmes de gestion reposaient sur une bonne compréhension fondamentale de la dynamique des populations de b. tabaci et tables de survie ont été une technique clée qui ont permis à cette compréhension. En Arizona, les tables de survie ont permis l’estimation et l’identification des forces importantes de mortalité de b. tabaci dans plusieurs cultures systèmes13,26, ont permis à la mesure de la dynamique de la mortalité comparativement à stratégies de gestion, y compris les effets non ciblés des insecticides14, ont fourni un moyen d’estimer les effets potentiels de non-cible fonctionnelles du coton transgénique produisant des protéines insecticides27, ont pris en charge rigoureuses évaluation d’une lutte biologique classique programme28 (Naranjo, données inédites) et aidé à explorer les effets comparatifs de top-down et bottom-up effets ravageurs dynamique29. Toutes ces applications ont déployé la méthodologie décrite ici. La méthode pourrait être utile pour l’étude de l’écologie des populations d’insectes dans un certain nombre de systèmes naturels et aménagés.

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methodology for developing life tables for sessile insects in the field using the whitefly, bemisia tabaci, in cotton as a model system

Tables de mortalité fournissent un moyen de mesurer les listes de naissance et de décès des populations au fil du temps. Ils ont aussi permet de quantifier les sources et les taux de mortalité dans les populations, qui a une variété d’applications en écologie, y compris les écosystèmes agricoles. Horizontale, ou fondées sur une cohorte, tableaux fournissent pour la méthode plus directe et précise de quantifier les taux vitaux de la population parce qu’elles suivent un groupe d’individus dans une population de la naissance à la mort à la vie. Ici, les protocoles sont présentés pour réaliser et analyser les tableaux de vie fondées sur une cohorte dans le domaine qui tire parti de la nature sessile des stades immatures de la vie d’un insecte nuisible planétaire, Bemisia tabaci. Les insectes sont situés sur la face inférieure des feuilles de coton et sont marquées par un petit cercle autour de l’insecte de dessin avec un stylo non toxique. Cet insecte peut alors être observé à plusieurs reprises au fil du temps à l’aide de loupes sur mesure développement d’un stade à l’autre et d’identifier les causes spécifiques au stade de la mort associée à la mortalité naturelle et introduit des forces. Les analyses expliquent comment mesurer correctement mortalité multiples forces cette loi en même temps au sein de chaque étape et comment utiliser ces données pour fournir les mesures dynamiques de population significative. La méthode ne tient pas directement compte de survie des adultes et de la reproduction, qui limite la déduction à la dynamique des stades immatures. Un exemple est présenté qui portait sur la mesure de l’impact du bas vers le haut (qualité de l’usine) et descendante (ennemis naturels) effets sur la dynamique de la mortalité de b. tabaci dans le système de coton.

Une cohorte de l’exemple est présentée dans le tableau 2 pour afficher une présentation typique et le calcul des résultats de tableau de la vie. Les données plus utiles sont capturées dans les taux de mortalité marginale de chaque facteur au sein de chaque étape. En convertissant ces taux en valeurs de k (protocole section 6), mortalité par étape au cours de tous les facteurs et facteur de mortalité sur toutes les étapes peuvent être facilement estimés, co...

Watch this Scientific Journal Video about Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System at JoVE.com

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Tables de survie permettent de quantifier les sources et les taux de mortalité dans les populations d’insectes et de contribuent à comprendre, de prédire et de manipuler la dynamique des populations dans les agroécosystèmes. Méthodes pour réaliser et analyser les tableaux de vie fondées sur une cohorte dans le domaine pour un insecte avec les stades immatures sessiles sont présentées.

Méthodologie pour l’élaboration des Tables de survie pour les insectes sessiles dans le champ à l’aide de l’aleurode, Bemisia tabaci, en coton comme modèle d’un système

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and ...

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Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Survival Analysis

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11.7K vues.  USDA-ARS, Arid-Land Agricultural Research Center. Tables de survie permettent de quantifier les sources et les taux de mortalité dans les populations d’insectes et de contribuent à comprendre, de prédire et de manipuler la dynamique des populations dans les agroécosystèmes. Méthodes pour réaliser et analyser les tableaux de vie fondées sur une cohorte dans le domaine pour un insecte avec les stades immatures sessiles sont présentées.

Vidéo: Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail millet. Recently, the 510 Mb genome of foxtail millet, S. italica, has been sequenced 1,2 and a 25x coverage genome sequence of the weedy relative S. viridis is in progress. S. viridis has a number of characteristics that make it a potentially excellent model genetic system including a rapid generation time, small stature, simple growth requirements, prolific seed production 3 and developed systems for both transient and stable transformation 4. However, the genetics of S. viridis is largely unexplored, in part, due to the lack of detailed methods for performing crosses. To date, no standard protocol has been adopted that will permit rapid production of seeds from controlled crosses.
The protocol presented here is optimized for performing genetic crosses in S. viridis, accession A10.1. We have employed a simple heat treatment with warm water for emasculation after pruning the panicle to retain 20-30 florets and labeling of flowers to eliminate seeds resulting from newly developed flowers after emasculation. After testing a series of heat treatments at permissive temperatures and varying the duration of dipping, we have established an optimum temperature and time range of 48 &deg;C for 3-6 min. By using this method, a minimum of 15 crosses can be performed by a single worker per day and an average of 3-5 outcross progeny per panicle can be recovered. Therefore, an average of 45-75 outcross progeny can be produced by one person in a single day. Broad implementation of this technique will facilitate the development of recombinant inbred line populations of S. viridis X S. viridis or S. viridis X S. italica, mapping mutations through bulk segregant analysis and creating higher order mutants for genetic analysis.

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

We have developed a methodology for performing crosses in Setaria viridis (S. viridis). The method involves pruning the panicle prior to a hot water treatment to kill viable pollen. Crosses are performed following a well-controlled growth regime and typically result in the recovery of 1 to 7 cross-pollinated seed/s per panicle.

Méthodes pour effectuer des croix dans<em&gt; Setaria viridis</em&gt;, Un nouveau système modèle pour les graminées

Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail ...

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Environnement

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

Un système d&#39;exposition accréditives pour l&#39;évaluation de suspension Sédiments effets sur la vie aquatique

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Controlled-release of Chlorine Dioxide in a Perforated Packaging System to Extend the Storage Life and Improve the Safety of Grape Tomatoes

A controlled-release chlorine dioxide (ClO2) pouch was developed by sealing a slurry form of ClO2 into semipermeable polymer film; the release properties of the pouch were monitored in containers with or without fruit. The pouch was affixed to the inside of a perforated clamshell containing grape tomatoes, and the effect on microbial population, firmness, and weight loss was evaluated during a 14 day storage period at 20 &#176;C. Within 3 days, the ClO2 concentration in the clamshells reached 3.5 ppm and remained constant until day 10. Thereafter, it decreased to 2 ppm by day 14. The ClO2 pouch exhibited strong antimicrobial activity, reducing Escherichia coli populations by 3.08 log CFU/g and Alternaria alternata populations by 2.85 log CFU/g after 14 days of storage. The ClO2 treatment also reduced softening and weight loss and extended the overall shelf life of the tomatoes. Our results suggest that ClO2 treatment is useful for extending the shelf life and improving the microbial safety of tomatoes during storage without impairing their quality.

Here, we describe a protocol for the application of a novel, slow-release ClO2 product that reduces spoilage and extends the shelf life of fresh fruit. The slow-release ClO2 product was added to standard commercial grape tomato packaging and tested against Escherichia coli and Alternaria alternata.

À libération contrôlée de dioxyde de chlore dans un système d&#39;emballage perforé pour prolonger la vie de stockage et améliorer la sécurité des tomates raisin

A controlled-release chlorine dioxide (ClO2) pouch was developed by sealing a slurry form of ClO2 into semipermeable polymer film; the release properties ...

Techniques for Investigating the Anatomy of the Ant Visual System

This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye anatomy of insects. These include traditional histological techniques optimized for work on ant eyes and adapted to work in concert with other techniques such as transmission electron microscopy (TEM) and scanning electron microscopy (SEM). These techniques, although vastly useful, can be difficult for the novice microscopist, so great emphasis has been placed in this article on troubleshooting and optimization for different specimens. We provide information on imaging techniques for the entire specimen (photo-microscopy and SEM) and discuss their advantages and disadvantages. We highlight the technique used in determining lens diameters for the entire eye and discuss new techniques for improvement. Lastly, we discuss techniques involved in preparing samples for LM and TEM, sectioning, staining, and imaging these samples. We discuss the hurdles that one might come across when preparing samples and how best to navigate around them.

This article outlines a suite of techniques in light and electron microscopy to study the internal and external eye anatomy of insects. These include several traditional techniques optimized for work on ant eyes, detailed troubleshooting, and suggestions for optimization for different specimens and regions of interest.

Techniques pour étudier l’anatomie du système visuel Ant

This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye ...

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, and the majority of extant heteropterans retain this trophic strategy. Some heteropterans have transitioned to feeding on vertebrate blood (such as the kissing bugs, Triatominae; and bed bugs, Cimicidae) while others have reverted to feeding on plants (most Pentatomomorpha). However, with the exception of saliva used by kissing bugs to facilitate blood-feeding, little is known about heteropteran venoms compared to the venoms of spiders, scorpions and snakes.
One obstacle to the characterization of heteropteran venom toxins is the structure and function of the venom/labial glands, which are both morphologically complex and perform multiple biological roles (defense, prey capture, and extra-oral digestion). In this article, we describe three methods we have successfully used to collect heteropteran venoms. First, we present electrostimulation as a convenient way to collect venom that is often lethal when injected into prey animals, and which obviates contamination by glandular tissue. Second, we show that gentle harassment of animals is sufficient to produce venom extrusion from the proboscis and/or venom spitting in some groups of heteropterans. Third, we describe methods to harvest venom toxins by dissection of anaesthetized animals to obtain the venom glands. This method is complementary to other methods, as it may allow harvesting of toxins from taxa in which electrostimulation and harassment are ineffective. These protocols will enable researchers to harvest toxins from heteropteran insects for structure-function characterization and possible applications in medicine and agriculture.

Although many insects in the suborder Heteroptera (Insecta: Hemiptera) are venomous, their venom composition and the functions of their venom toxins are mostly unknown. This protocol describes methods to harvest heteropteran venoms for further characterization, using electrostimulation, harassment, and gland dissection.

Récolte de venin toxines d’Assassin Bugs et autres insectes hétéroptères

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, ...

A Precise and Autonomous System for the Detection of Insect Emergence Patterns

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of emerging insects they can detect. In order to obtain precise measurement of insect emergence, it is necessary for systems to be semi-automated and able to measure large numbers of emerging insects. We addressed these issues by designing and building a system that is automated and can measure emergence of up to 1200 insects. We modified the existing &#34;falling-ball&#34; system using Arduino microcontrollers to automate data collection and expand the sample size through multiple data channels. Multiple data channels enable the user to not only increase their sample size, but also allows for multiple treatments to be run simultaneously in a single experiment. Furthermore, we created an R script to automatically visualize the data as a bubble plot, while also calculating the median day and time of emergence. The current system was designed using 3D printing so the user can modify the system to be adjusted for different species of insects. The goal of this protocol is to investigate important questions in chronobiology and stress physiology, using this precise and automated system to measure insect emergence patterns.

Measurement of insect emergence patterns requires precision. Existing systems are only semi-automated and sample size is limited. We addressed these issues by designing a system using microcontrollers to precisely measure the time of emergence of large numbers of emerging insects.

Un système précis et autonome pour la détection des patrons d’émergence insectes

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

The efficacy of public health pesticides targeting nuisance and disease-vector insects is not uniform across different ecological zones. Here we present a system of techniques using captive vector insects as sentinels for pesticide efficacy to derive electronic maps supporting the standard evaluation of pesticides across multiple environments.

Visualisation de l’efficacité des Pesticides contre les moustiques vecteur de maladie dans le domaine

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

Un test d’Amplification isotherme (lampe) médiée par la boucle pour une Identification rapide de Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

An Induction System for Clustered Stomata by Sugar Solution Immersion Treatment in Arabidopsis thaliana Seedlings

Stomatal movement mediates plant gas exchange, which is essential for photosynthesis and transpiration. Stomatal opening and closing are accomplished by a significant increase and decrease in guard cell volume, respectively. Because shuttle transport of ions and water occurs between guard cells and larger neighboring epidermal cells during stomatal movement, the spaced distribution of plant stomata is considered an optimal distribution for stomatal movement. Experimental systems for perturbing the spaced pattern of stomata are useful to examine the spacing pattern's significance. Several key genes associated with the spaced stomatal distribution have been identified, and clustered stomata can be experimentally induced by altering these genes. Alternatively, clustered stomata can be also induced by exogenous treatments without genetic modification. In this article, we describe a simple induction system for clustered stomata in Arabidopsis thaliana seedlings by immersion treatment with a sucrose-containing medium solution. Our method is easy and directly applicable to transgenic or mutant lines. Larger chloroplasts are presented as a cell biological hallmark of sucrose-induced clustered guard cells. In addition, a representative confocal microscopic image of cortical microtubules is shown as an example of intracellular observation of clustered guard cells. The radial orientation of cortical microtubules is maintained in clustered guard cells as in spaced guard cells in control conditions.

An Induction System for Clustered Stomata by Sugar Solution Immersion Treatment in Arabidopsis thaliana Seedlings

The goal of this protocol is to demonstrate how to induce clustered stomata in cotyledons of Arabidopsis thaliana seedlings by immersion treatment with a sugar-containing medium solution and how to observe intracellular structures such as chloroplasts and microtubules in the clustered guard cells using confocal laser microscopy.

Un système d’Induction de stomates groupées par traitement de Immersion de Solution de sucre chez les plantules de Arabidopsis thaliana

Stomatal movement mediates plant gas exchange, which is essential for photosynthesis and transpiration. Stomatal opening and closing are accomplished by a ...

A Gnotobiotic System for Studying Microbiome Assembly in the Phyllosphere and in Vegetable Fermentation

The phyllosphere, the above ground portion of the plant that can be colonized by microbes, is a useful model system to identify processes of microbial community assembly. This protocol outlines a system for studying microbial community dynamics in the phyllosphere of Napa cabbage plants. It describes how to grow germ-free plants in test tubes with a calcined clay and nutrient broth substrate. Inoculation of germ-free plants with specific microbial cultures provides opportunities to measure microbial growth and community dynamics in the phyllosphere. Through the use of sterile vegetable extract produced from cabbages shifts in microbial communities that occur during fermentation can also be assessed. This system is relatively simple and inexpensive to set up in the lab and can be used to address key ecological questions in microbial community assembly. It also provides opportunities to understand how phyllosphere community composition can impact the microbial diversity and quality of vegetable fermentations. This approach for developing gnotobiotic cabbage phyllosphere communities could be applied to other wild and agricultural plant species.

A method of growing germ-free Napa cabbages has been developed which enables researchers to evaluate how single microbial species or multispecies microbial communities interact on cabbage leaf surfaces. A sterile vegetable extract is also presented which can be used to measure shifts in community composition during vegetable fermentation.

Un système gnotobiotique pour l’étude de l’assemblage de microbiome dans la Phyllosphère et dans la fermentation végétale

The phyllosphere, the above ground portion of the plant that can be colonized by microbes, is a useful model system to identify processes of microbial ...